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Analysis of Genes and Genomes

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  • Àú : Reece, Richard
  • ÃâÆÇ»ç : Wiley
  • ¹ßÇà : 2004³â 01¿ù 01ÀÏ
  • Âʼö : 0
  • ISBN : 9780470843802
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Prefacep. xiii
Acknowledgementsp. xv
Abbreviations and acronymsp. xvii
DNA: Structure and functionp. 1
Nucleic acid is the material of heredityp. 2
Structure of nucleic acidsp. 7
The double helixp. 11
The antiparallel helixp. 12
Base pairs and stackingp. 14
Gaining access to information with the double helix without breaking it apartp. 16
Hydrogen bondingp. 17
Reversible denaturing of DNAp. 18
Structure of DNA in the cellp. 21
The eukaryotic nucleosomep. 24
The replication of DNAp. 28
DNA polymerasesp. 31
The replication processp. 33
Recombinationp. 37
Genes and genomesp. 39
Genes within a genomep. 40
Transcriptionp. 43
Transcription in prokaryotesp. 43
Transcription in eukaryotesp. 46
RNA processingp. 54
RNA splicingp. 55
Alternative splicingp. 58
Translationp. 59
Basic techniques in gene analysisp. 65
Restriction enzymesp. 66
Types of restriction-modification systemp. 70
Other modification systemsp. 72
How do type II restriction enzymes work?p. 74
Joining DNA moleculesp. 76
The basics of cloningp. 78
Bacterial transformationp. 84
Chemical transformationp. 86
Electroporationp. 87
Gene gunp. 88
Gel electrophoresisp. 88
Polyacrylamide gelsp. 89
Agarose gelsp. 89
Pulsed-field gel electrophoresisp. 95
Nucleic acid blottingp. 98
Southern blottingp. 100
The compass points of blottingp. 102
DNA purificationp. 103
Vectorsp. 109
Plasmidsp. 112
pBR322p. 116
pUC plasmidsp. 119
Selectable markersp. 122
[lambda] vectorsp. 126
Cosmid vectorsp. 135
M13 vectorsp. 137
Phagemidsp. 140
Artificial chromosomesp. 142
YACsp. 143
PACsp. 146
BACsp. 148
HACsp. 149
Polymerase chain reactionp. 153
PCR reaction conditionsp. 159
Thermostable DNA polymerasesp. 162
Template DNAp. 164
Oligonucleotide primersp. 165
Synthesis of oligonucleotide primersp. 167
Primer mismatchesp. 169
PCR in the diagnosis of genetic diseasep. 173
Cloning PCR productsp. 175
RT-PCRp. 177
Real-time PCRp. 179
Applications of PCRp. 181
Cloning a genep. 183
Genomic librariesp. 185
cDNA librariesp. 191
Directional cDNA cloningp. 196
PCR based librariesp. 199
Subtraction librariesp. 200
Library construction in the post-genome erap. 204
Gene identificationp. 205
Screening by nucleic acid hybridizationp. 206
Immunoscreeningp. 211
Screening by functionp. 216
Screening by interactionp. 217
Phage displayp. 218
Two-hybrid screeningp. 218
Problems, and some solutions, with two-hybrid screeningp. 225
Other interaction screens--variations on a themep. 228
One hybridp. 229
Three hybridp. 229
Reverse two hybridp. 229
Creating mutationsp. 231
Creating specific DNA changes using primer extension mutagenesisp. 233
Strand selection methodsp. 237
Phosphorothioate strand selectionp. 237
dut[superscript -] ung[superscript -] (or Kunkel) strand selectionp. 238
Cassette mutagenesisp. 240
PCR based mutagenesisp. 241
QuikChange mutagenesisp. 248
Creating random mutations in specific genesp. 250
Protein engineeringp. 254
Protein production and purificationp. 257
Expression in E. colip. 258
The lac promoterp. 259
The tac promoterp. 259
The [lambda]P[subscript L] promoterp. 260
The T7 expression systemp. 261
Expression in yeastp. 265
Saccharomyces cerevisiaep. 265
The GAL systemp. 266
The CUP1 systemp. 268
Pichia pastorisp. 268
Schizosaccharomyces pombep. 269
Expression in insect cellsp. 269
Expression in higher-Eukaryotic cellsp. 272
Tet-on/Tet-off systemp. 272
Protein purificationp. 275
The His-tagp. 276
The GST-tagp. 279
The MBP-tagp. 282
IMPACTp. 282
TAP-taggingp. 286
Genome sequencing projectsp. 287
Genomic mappingp. 289
Genetic mappingp. 290
Physical mappingp. 293
Nucleotide sequencingp. 295
Manual DNA sequencingp. 296
Automated DNA sequencingp. 300
Genome sequencingp. 303
The human genome projectp. 305
Finding genesp. 307
Gene assignmentp. 309
Bioinformaticsp. 311
Post-genome analysisp. 313
Global changes in gene expressionp. 314
Differential displayp. 315
Microarraysp. 317
ChIPs with everythingp. 324
Protein function on a genome-wide scalep. 327
Knock-out analysisp. 327
Antisense and RNA interference (RNAi)p. 329
Genome-wide two-hybrid screensp. 333
Protein detection arraysp. 335
Structural genomicsp. 335
Engineering plantsp. 341
Cloning in plantsp. 341
Agrobacterium tumefaciensp. 342
Direct nuclear transformationp. 347
Viral vectorsp. 348
Chloroplast transformationp. 350
Commercial exploitation of plant transgenicsp. 354
Delayed ripeningp. 354
Insecticidal resistancep. 355
Herbicidal resistancep. 356
Viral resistancep. 357
Fungal resistancep. 358
Terminator technologyp. 358
Ethics of genetically engineered cropsp. 360
Engineering animal cellsp. 361
Cell culturep. 361
Transfection of animal cellsp. 362
Chemical transfectionp. 363
Electroporationp. 364
Liposome-mediated transfectionp. 364
Peptidesp. 366
Direct DNA transferp. 366
Viruses as vectorsp. 367
SV40p. 367
Adenovirusp. 369
Adeno-associated virus (AAV)p. 371
Retrovirusp. 372
Selectable markers and gene amplification in animal cellsp. 375
Expressing genes in animal cellsp. 378
Engineering animalsp. 379
Pronuclear injectionp. 381
Embryonic stem cellsp. 384
Nuclear transferp. 390
Gene therapyp. 396
Examples and potential of gene therapyp. 398
Glossaryp. 401
Proteinsp. 409
p. 409
p. 410
p. 411
Nobel prize winnersp. 413
Referencesp. 417
Indexp. 459
Table of Contents provided by Rittenhouse. All Rights Reserved.

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"This beautifully illustrated textbook provides a clear guide to the tools and techniques of genetic engineering, gene cloning and molecular biology. All aspects of genetic engineering in the post-genomic era are covered, beginning with the basics of DNA structure and DNA metabolism. Using an example-driven approach, the fundamentals of creating mutations in DNA, cloning in bacteria, yeast, plants and animals are all clearly presented. Strong emphasis is placed on the latest, post genomic technologies including DNA macro and microarrays, genome-wide two hybrid analysis, proteomics and bioinformatics. A modern post-genome era introduction to key techniques used in genetic engineering. An example driven past-to-present approach to allow the experiments of today to be placed in an historical context The book is beautifully illustrated in full-colour throughout. Associated website including updates, additional content and illusions "

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